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. 2020 Jan 16;9:e52056. doi: 10.7554/eLife.52056

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© 2020, Zhou et al

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Figure 4. — (A) Western blots of myelin proteins in the cerebral cortex (CTX), the hippocampus (Hip), the cerebellum (CB), spinal cord, the corpus callosum (CC), and optic nerve (ON) from Gab1^f/+;Olig1-cre (ctrl1) vs. Gab1^f/f;Olig1-cre (KO1) mice or Gab1^f/f (ctrl2) vs. Gab1^f/f;Cspg2-cre (KO2) mice at P21. (B) MBP expression was normalized to GAPDH and percentage changes are shown. In the order of ctrl1, KO1, ctrl2, and KO2, changes are: CTX, 100 ± 2% and 20 ± 8% (p=0.000030), 100 ± 2% and 51 ± 5% (p=0.000051); Hip, 100 ± 2% and 46 ± 2% (p=0.000001), 100 ± 3% and 18 ± 4% (p=0.000001); CB, 100 ± 3% and 38 ± 6% (p=0.000034), 100 ± 2% and 84 ± 3%(p=0.0058); spinal cord, 100 ± 3% and 47 ± 3% (p=0.000007), 100 ± 3% and 76 ± 4% (p=0.0013); CC, 100 ± 3% and 32 ± 3% (p=0.000002), 100 ± 3% and 52 ± 7%(p=0.00036); ON, 100 ± 3% and 42 ± 4% (p=0.000014), 100 ± 3% and 33 ± 5% (p=0.000017). n = 4/group. t-test, df = t(7). (C) Myelin proteins in myelin fractions of corpus callosum from Gab1^f/+;Olig1-cre and Gab1^f/f;Olig1-cre mice at P60. GAPDH was internal control. Expressions of MBP, PLP and MOG were normalized to corresponding GAPDH and the percentage changes are shown. MBP: 100 ± 4% (Gab1^f/+;Olig1-cre) and 23 ± 6% (Gab1^f/f;Olig1-cre) (p=0.000023). PLP: 100 ± 4% (Gab1^f/+;Olig1-cre) and 18 ± 7% (Gab1^f/f;Olig1-cre) (p=0.000039). MOG: 100 ± 5% (Gab1^f/+;Olig1-cre) and 45 ± 10% (Gab1^f/f;Olig1-cre) (p=0.0011). n = 4/group, t-test, df = t(7). (D) Western blots of myelin proteins in the cerebral cortex (CTX) and the cerebellum (CB) from WT, Gab2^-/- and Gab2^+/- mice at P21. The experiment was repeated four times. Gray dots indicate individual data points. ***p<0.01.

Figure 4—figure supplement 1. — (A) Western blots of myelin proteins in the cerebral cortex (CTX), the hippocampus (Hip), the cerebellum (CB), spinal cord, the corpus callosum (CC), and optic nerve (ON) from Gab1^f/+;Olig1-cre (ctrl1) vs. Gab1^f/f;Olig1-cre (KO1) mice or Gab1^f/f (ctrl2) vs. Gab1^f/f;Cspg2-cre (KO2) mice at P21. (B) MBP expression was normalized to GAPDH and percentage changes are shown. In the order of ctrl1, KO1, ctrl2, and KO2, changes are: CTX, 100 ± 2% and 20 ± 8% (p=0.000030), 100 ± 2% and 51 ± 5% (p=0.000051); Hip, 100 ± 2% and 46 ± 2% (p=0.000001), 100 ± 3% and 18 ± 4% (p=0.000001); CB, 100 ± 3% and 38 ± 6% (p=0.000034), 100 ± 2% and 84 ± 3%(p=0.0058); spinal cord, 100 ± 3% and 47 ± 3% (p=0.000007), 100 ± 3% and 76 ± 4% (p=0.0013); CC, 100 ± 3% and 32 ± 3% (p=0.000002), 100 ± 3% and 52 ± 7%(p=0.00036); ON, 100 ± 3% and 42 ± 4% (p=0.000014), 100 ± 3% and 33 ± 5% (p=0.000017). n = 4/group. t-test, df = t(7). (C) Myelin proteins in myelin fractions of corpus callosum from Gab1^f/+;Olig1-cre and Gab1^f/f;Olig1-cre mice at P60. GAPDH was internal control. Expressions of MBP, PLP and MOG were normalized to corresponding GAPDH and the percentage changes are shown. MBP: 100 ± 4% (Gab1^f/+;Olig1-cre) and 23 ± 6% (Gab1^f/f;Olig1-cre) (p=0.000023). PLP: 100 ± 4% (Gab1^f/+;Olig1-cre) and 18 ± 7% (Gab1^f/f;Olig1-cre) (p=0.000039). MOG: 100 ± 5% (Gab1^f/+;Olig1-cre) and 45 ± 10% (Gab1^f/f;Olig1-cre) (p=0.0011). n = 4/group, t-test, df = t(7). (D) Western blots of myelin proteins in the cerebral cortex (CTX) and the cerebellum (CB) from WT, Gab2^-/- and Gab2^+/- mice at P21. The experiment was repeated four times. Gray dots indicate individual data points. ***p<0.01.